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MicroRNA-21 preserves the fibrotic mechanical memory of mesenchymal stem cells

Nature Materials volume 16, pages 379–389 (2017)Cite this article

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Abstract

Expansion on stiff culture substrates activates pro-fibrotic cell programs that are retained by mechanical memory. Here, we show that priming on physiologically soft silicone substrates suppresses fibrogenesis and desensitizes mesenchymal stem cells (MSCs) against subsequent mechanical activation in vitro and in vivo, and identify the microRNA miR-21 as a long-term memory keeper of the fibrogenic program in MSCs. During stiff priming, miR-21 levels were gradually increased by continued regulation through the acutely mechanosensitive myocardin-related transcription factor-A (MRTF-A/MLK-1) and remained high over 2 weeks after removal of the mechanical stimulus. Knocking down miR-21 once by the end of the stiff-priming period was sufficient to erase the mechanical memory and sensitize MSCs to subsequent exposure to soft substrates. Soft priming and erasing mechanical memory following cell culture expansion protects MSCs from fibrogenesis in the host wound environment and increases the chances for success of MSC therapy in tissue-repair applications.

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Figure 1: MSC isolation and expansion on pathophysiologically stiff/soft substrates.
Figure 2: MSCs undergo fibrotic mechanical priming on pathophysiologically stiff substrates.
Figure 3: MRTF-A regulates miR-21 transcription.
Figure 4: MiR-21 controls fibrogenesis of MSCs at the gene transcription level.
Figure 5: MiR-21 preserves mechanical memory in MSCs.
Figure 6: Transplantation of soft-primed MSCs reduces scarring by suppressing myofibroblast formation and tissue contracture.

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Acknowledgements

We are grateful to J. Lammerding (Cornell University, Ithaca, New York, USA) and M. K. Vartiainen (University of Helsinki, Helsinki, Finland) for providing full-length and truncated constructs of MRTF-A-GFP. This research was supported by grants to B.H. from the Canadian Institutes of Health Research CIHR (grants no. 210820, no. 286920 and no. 286720), the Collaborative Health Research Programme (CIHR/NSERC grants no. 1004005 and no. 413783), and the Canada Foundation for Innovation and Ontario Research Fund (CFI/ORF grant no. 26653) and grants to A.Kapus from the CIHR (MOP-106625 and 130463), the Canadian Foundation of Innovation and the Kidney Foundation of Canada. Data presented herein were further funded from the European Union’s Seventh Framework Program (FP7/2007-2013) under grant agreement no. 237946 and the CIHR Cell Signals Training programme. C.L. was supported by the CIHR Training Program in Regenerative Medicine (TPRM) and an Ontario graduate fellowship.

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  1. Chen Xi Li, Nilesh P. Talele and Stellar Boo: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Tissue Repair and Regeneration, Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Toronto, Ontario M5S 3E2, Canada

    Chen Xi Li, Nilesh P. Talele, Stellar Boo, Anne Koehler, Ericka Knee-Walden & Boris Hinz

  2. Draper, Cambridge, Massachusetts 02139, USA

    Jenna L. Balestrini

  3. and Department of Surgery, Keenan Research Centre in the Li Ka Shing Knowledge Institute in the St. Michael’s Hospital, University of Toronto, Toronto, Ontario M5B 1W8, Canada

    Pam Speight & Andras Kapus

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Contributions

C.X.L., B.H., N.P.T. and J.L.B. conceived the ideas and designed the experiments. C.X.L., E.K.-W., N.P.T., P.S., A.Koehler and S.B. performed experiments and analysed the data. C.X.L., B.H., N.P.T., A.Kapus and J.L.B. interpreted the data and wrote the manuscript. B.H. directed the work.

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Correspondence to Boris Hinz.

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Li, C., Talele, N., Boo, S. et al. MicroRNA-21 preserves the fibrotic mechanical memory of mesenchymal stem cells. Nature Mater 16, 379–389 (2017). https://doi.org/10.1038/nmat4780

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